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小麦条锈菌(Puccinia striiformis f.sp.tritici)是专性程度很高的活体寄生菌,通过分子技术对其进行群体遗传结构研究的过程中,基因组DNA提取是基础之一。本研究利用Chelex-100法和CTAB法直接提取罹条锈病的小麦病叶片上的小麦条锈病菌的基因组DNA,使用PCR对所得到的DNA进行真菌核糖体rDNA-ITS和小麦条锈病菌特异性引物CYR33进行检测,对2种基因组DNA的提取方法进行比较分析,结果表明Chel-ex-100法和CTAB法都能够适合于从罹病组织中直接提取小麦条锈病菌的基因组DNA,但是Chelex-100法较CTAB法所需的样品量少,操作步骤少,操作简单,并且整个过程无需使用有机溶剂进行抽提,对人体比较安全。通过PCR检测DNA提取情况,发现Chelex-100法提取的DNA的PCR结果优于CTAB法,且能够满足基于PCR的群体遗传结构的分析等研究。
Puccinia striiformis f. Sp. Tritici is a highly obligate living parasite. Genomic DNA extraction is one of the basic steps in the study of population genetic structure by molecular techniques. In this study, Chelex-100 method and CTAB method were used to directly extract the genomic DNA of wheat stripe rust on the stripe rust-resistant wheat leaves. PCR was used to analyze the specific DNA of the rDNA-ITS and wheat stripe rust The primers CYR33 were used to detect the genomic DNA extracted from two genomic DNAs. The results showed that both the Chel-ex-100 and CTAB methods were suitable for the direct extraction of genomic DNA from diseased tissues of the stripe rust, but Chelex-100 Compared with the CTAB method, the method requires less samples, less operation steps and simple operation, and the whole process does not need to use organic solvents for extraction, which is safer to the human body. The results of DNA extraction by PCR showed that the PCR results of Chelex-100 method were superior to those of CTAB method and could be used to analyze the genetic structure of PCR-based population.